建立了利用高效液相色谱同时测定粪产碱杆菌菌液中5种有机酸代谢产物的分析方法。采用Agilent Eclipse XDB-C18色谱柱(4.6×150 mm,5 μm),流动相为V(磷酸氢二钠):V(甲醇)=97:3,流速0.6 mL/min,检测波长210 nm,柱温35℃,进样量20 μL。结果表明,该方法在12 min内简便快速地分离出5种有机酸(丁二酸、乳酸、柠檬酸、乙醛酸和苹果酸),且峰形良好、分离度好,线性范围内各酸的浓度与色谱峰面积的线性关系良好,相关系数不低于0.999 5,加标回收率为99.22%~103.60%,相对标准偏差小于5%。该方法简便快速,具有前处理简单、分离效果好、精密度高、回收率高等特点,可用于同时测定菌液中5种有机酸的含量。

An analytical method for simultaneous determination of five kinds of organic acid metabolites in Alcaligenes faecalis is established by means of high performance liquid chromatography.An Agilent Eclipse XDB-C18 column (4.6×150 mm,5 μm) is used,the mixed mobile phase is composed of disodium hydrogen phosphate:methanol=97:3,the flow rate is 0.6 mL·min^-1,the detection wavelength is 210 nm,the column temperature is 35℃,and injection volume is 20 μL.Results show that this method can easily and quickly separate five kinds of organic acids (succinic acid,lactic acid,citric acid,glyoxylic acid and malic acid) within 10 minutes,shows good peak shape and good resolution,and exhibits good linear relationship between the concentrations of each acid and chromatographic peak area.It delivers a correlation coefficient not less than 0.999 5,a standard addition recovery rate of 99.22%-103.60%,and a relative standard deviation less than 5%.It is verified that this method is simple and rapid,and shows the characteristics such as simple pretreatment,good separation effect,high precision and high recovery rate.It can be used to simultaneously determine the contents of five kinds of organic acids.

万欣(1998-),男,硕士生